title: Ovarian cancer ascites-derived vitronectin and fibronectin: combined purification, molecular features and effects on cell response.
abstract: BACKGROUND
  Intra-abdominal ascites is a complication of ovarian cancers and constitutes a permissive microenvironment for metastasis. Since fibronectin and vitronectin are key actors in ovarian cancer progression, we investigated their occurrence and molecular characteristics in various ascites fluids and the influence of these ascites-derived proteins on cell behavior.
  
  
  METHODS
  Fibronectin and vitronectin were investigated by immunoblotting within various ascites fluids. A combined affinity-based protocol was developed to purify both proteins from the same sample. Each purified protein was characterized with regard to its molecular features (molecular mass of isoforms, tryptophan intramolecular environment, hydrodynamic radii), and its influence on cell adhesion.
  
  
  RESULTS
  Fibronectin and vitronectin were found in all tested ascites. Several milligrams of purified proteins were obtained from ascites of varying initial volumes. Molecular mass isoforms and conformational lability of proteins differed according to the ascites of origin. When incorporated into the cancer cell environment, ascites-derived fibronectin and vitronectin supported cell adhesion and migration with various degrees of efficiency, and induced the recruitment of integrins into focal contacts.
  
  
  CONCLUSIONS
  To our knowledge, this is the first combined purification of two extracellular matrix proteins from a single pathological sample containing a great variety of bioactive molecules. This study highlights that ascites-derived fibronectin and vitronectin exhibit different properties depending on the ascites.
  
  
  CONCLUSIONS
  Investigating the relationships between the molecular properties of ascites components and ovarian cancer cell phenotype according to the ascites may be critical for a better understanding of the recurrence of this lethal disease and for further biomarker identification.
authors.affiliation: Institut des Matériaux, Université de Cergy-Pontoise, Cergy-Pontoise Cedex, France.
authors.forename: Ludovic
authors.initials: L
authors.lastname: Carduner
authors.forename: Rémy
authors.initials: R
authors.lastname: Agniel
authors.forename: Sabrina
authors.initials: S
authors.lastname: Kellouche
authors.forename: Cédric R
authors.initials: CR
authors.lastname: Picot
authors.forename: Cécile
authors.initials: C
authors.lastname: Blanc-Fournier
authors.forename: Johanne
authors.initials: J
authors.lastname: Leroy-Dudal
authors.forename: Franck
authors.initials: F
authors.lastname: Carreiras
mesh_terms: D001201:Ascites / Q000378:metabolism*; D005260:Female; D005353:Fibronectins / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D010051:Ovarian Neoplasms / Q000378:metabolism* / Q000473:pathology; D011487:Protein Conformation; D019096:Vitronectin / Q000737:chemistry / Q000378:metabolism*
chemical_list: D005353:Fibronectins; D019096:Vitronectin
keywords: Affinity chromatography; Ascites; ECM; Fibronectin; Fn; Microenvironment; Vitronectin; Vn; extracellular matrix; fibronectin; kDa; kilodaltons; vitronectin